Abstract
Specific, end-labeled DNA fragments can be simply and rapidly prepared using the polymerase chain reaction (PCR). Such fragments are suitable for use in DNase I protection footprint assays, chemical sequencing reactions, and for the production and analysis of paused RNA polymerase transcription complexes. Moreover, a general means of introducing a specific mutation at any position along the length of such PCR-generated fragments is described. These procedures, which can circumvent the need for large-scale phage or plasmid growths, preparative gel-electrophoresis and the screening of molecular clones, can facilitate the rapid study of sequence-specific interactions of proteins and DNA. A rapid means of removing excess oligonucleotide primers from completed PCRs is also described.
Keywords
BiologyDNAGeneticsNucleic acidMutagenesisLibrary scienceComputational biologyMutationGeneComputer science
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Publication Info
- Year
- 1988
- Type
- article
- Volume
- 16
- Issue
- 15
- Pages
- 7351-7367
- Citations
- 2394
- Access
- Closed
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Cite This
Russell Higuchi,
Barbara Krummel,
Randall K. Saiki
(1988).
A general method of<i>in vitro</i>preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions.
Nucleic Acids Research
, 16
(15)
, 7351-7367.
https://doi.org/10.1093/nar/16.15.7351
Identifiers
- DOI
- 10.1093/nar/16.15.7351